一般社団法人
日本細胞生物学会Japan Society for Cell Biology

4% PFA at 4oC over night

0.5% Triton in PBS

4% PFA at 4oC over night

0.5% Triton in PBS

4% PFA at 4oC over night

0.5% Triton in PBS

4%PFA (in phosphate buffer)

0.1% TritonX-100/PBS

3％PFA,　0.05%saponin

PC12細胞の免疫蛍光染色（3%PFA固定、0.05%saponinで膜のpermeabilize）よく染まった Good staining for IF of PC12 with 3%PFA, 0.05%saponin

3%PFA (in PBS)

MeOH

MeOH>TCA

IF: -20℃ methanol or acetone:methanol=1:1

IF: -20℃ methanol or acetone:methanol=1:1

IF: -20℃ methanol or acetone:methanol=1:1

チューブリンにも反応するので要注意
Also recognize tubulin

IF: -20℃ methanol or acetone:methanol=1:1

IF: -20℃ methanol or acetone:methanol=1:1

IF: -20℃ methanol or acetone:methanol=1:1

IF: -20℃ methanol or acetone:methanol=1:1

IF: -20℃ methanol or acetone:methanol=1:1

EEA1(mono)とパターン違う。
Staininig pattern is different from the one by EEA1 monoclonal Ab.

【PFA+Triton】:○【PFA条件】3%PFA/PBS　37℃10min【Triton条件】0.1%　Triton-X100/PBS

IFでBGあるが使える
Some background staining appears in IF staininig.

IFで使えない。
Not suitable for IF.

【PFA+Triton】:◎,【PFA+Saponin】:○【PFA条件】3%PFA/PBS　37℃10min【Triton条件】0.1%　Triton-X100/PBS【saponin条件】0.05%　saponin /PBS

MDCKII OK

4%PFA+0.1%Saponin :○

マウスのみ

PFA (fix cells by adding 1/10 vol of 37% formalin directly to the culture medium)+Triton

【3.7% formalin solution+Triton 0.2%, 5min】:○ formalin solution is made by dilution of 37% formalin (Nacalai Inc.) by PBS or Ringer's solution

WB:Blocking/3%BSA-TBS, 1stAb/3%BSA-TBS, 2ndAb/3% skin milk-TBSが良い。
ホルマリンは37％ホルマリン液（ナカライ）をPBSやRingerで１/10に希釈
Formalin solution is made by dilution of 37% formalin (Nacalai Inc.) by PBS or Ringer's solution

3%PFA (in PBS)

0.05%saponin

3%PFA in PBS

0.05% saponin

methanol でもOK
Fair staining by methanol fix.

3%PFA in PBS

0.05% saponin

3%PFA in PBS

0.05% saponin

3%PFA (in PBS)

0.05%saponin

included in Coated vesicle sampler kit

3%PFA (in PBS)

0.05%saponin

included in Coated vesicle sampler kit

3%PFA (in phosphate buffer)

3%PFA (in PBS)

0.05%saponin

4% PFA

Fof ELISA

1%PFA, 4%PFA, MeOH, TCA

【IS】:○

【PFA+Triton】:○,【PFA+Saponin】:○,【IS】:×,【MeOH】:○【PFA条件】3%PFA/PBS　37℃10min【MeOH条件】MeOH 100% -20℃ 5min【Triton条件】0.1%　Triton-X100/PBS【saponin条件】0.05%　saponin /PBS

Immunogen: Lumenal domain

human専用

4%PFA (in phosphate buffer)

4％PFA +0.1％Triton

4％PFA +0.1％Triton

4%PFA (in phosphate buffer)

4%PFA (in phosphate buffer)

4%PFA (in PBS)

PFA-fixed cryosection/HistoVT one (nacalai) 70˚C 20min

a type of pyramidal neuron

PFA (fix cells by adding 1/10 vol of 37% formalin directly to the culture medium)+Triton or PFA/GA + Triton or MeOH or TCA

4％PFA +0.1％Triton

4%PFA (in phosphate buffer)

0.1% TritonX-100/PBS

4 % PFA fix, PBS+0.05 %Tween20+ 0.5 % TSA block+1 % Donkey serum

1%PFA, 4%PFA, TCA

1%PFA, TCA

3%PFA (in PBS)

0.05%saponin

included in Coated vesicle sampler kit

3%PFA (in phosphate buffer)

【PFA+Triton】:○,【MeOH】:◎,【TCA】:○【PFA条件】3%PFA/PBS　37℃10min【MeOH条件】MeOH 100% -20℃ 5min【TCA条件】TCA 10%/H2O on ice 15min【Triton条件】0.1%　Triton-X100/PBS

ジギトニンでは見えない。MDCKIIダメ。
Digitonin is not suitable for permeabilization. Cannot stain MDCKII

Weakly cross-reactive with unknown protein (~46 kDa).

4% PFA/PBS

3%PFA in PBS

0.05% saponin

Ab in Chaperone sampler kit (BD Transduction Lab)

3%PFA in PBS

0.05% saponin

Ab in Chaperone sampler kit (BD Transduction Lab)

3%PFA (in PBS)

0.05%saponin

4％PFA +0.1％Triton

TCA, 4%PFA

2％ PFA 0.5%Triton, RT 15min

【抗原の種、部位】human 362-585 a.a.

3%PFA (in PBS)

4% PFA/PBS

3%PFA in PBS

0.05% saponin

3%PFA (in PBS)

通常の染色法で大丈夫
Stains well by ordinary staining procedures.

3%PFA (in PBS)

3%PFA (in PBS)

4%PFA/PBS

ice cold Methanol

4%PFA/PBS

signal decreases from 3h after oocyte activation

ice cold Methanol

signal decreases from 3h after oocyte activation

TCA>>4%PFA Ezrinに特異性はなくRadixin, Moesinも同様に認識。
Stains ezrin, radixin, and moesin.

3%PFA (in PBS)

0.05%saponin

3%PFA (in PBS)

0.05％saponin

4%PFA 大変弱い
Very weak staining

3%PFA (in PBS), 0.05% saponin

4％PFA +0.1％Triton

4％PFA +0.1％Triton

4％PFA +0.1％Triton

TCA>MeOH>1%PFA

交差する動物種：マウス
【FACS】:○（final conc）20 μg/mL

4％PFA +0.1％Triton

ダマができる

核小体が染まる、二次抗体　anti-mouse Alexa555及びanti-mouse Cy3がanti-FLAG (poly)を認識する可能性あり。要注意
Stains nucleoles.

IPは非常によい
Very good for IP

【3.7% formalin solution+0.5%Triton】:○ formalin solution is made by dilution of 37% formalin (Nacalai Inc.) by PBS or Ringer's solution

WB:Blocking/3%BSA-TBS, 1stAb/3%BSA-TBS, 2ndAb/3% skin milk-TBSが良い。
ホルマリンは37％ホルマリン液（ナカライ）をPBSやRingerで１/10に希釈
Formalin solution is made by dilution of 37% formalin (Nacalai Inc.) by PBS or Ringer's solution

【3.7% formalin solution+0.5%Triton】:○ formalin solution is made by dilution of 37% formalin (Nacalai Inc.) by PBS or Ringer's solution

WB:Blocking/3%BSA-TBS, 1stAb/3%BSA-TBS, 2ndAb/3% skin milk-TBSが良い。
ホルマリンは37％ホルマリン液（ナカライ）をPBSやRingerで１/10に希釈
Formalin solution is made by dilution of 37% formalin (Nacalai Inc.) by PBS or Ringer's solution

4％PFA +0.1％Triton

4%PFA (in PBS)

PFA-fixed cryosection/HistoVT one (nacalai) 70˚C 20min

CA1 pyramidal cell

methanol

3%PFA in PBSで染まらない
Not stained by 3%PFA in PBS fixation

3%PFA (in PBS), methanol

3%PFA (in PBS), methanol

4％PFA +0.1％Triton

3%PFA (in PBS)

3%PFA (in PBS)

cappelのものがベスト
Ab from Cappel is the best for beta-galactosidase antibody

3%PFA (in PBS)

cappelのものがベスト
Ab from Cappel is the best for beta-galactosidase antibody

3%PFA (in PBS)

cappelのものがベスト
Ab from Cappel is the best for beta-galactosidase antibody

【Formalin in Ringer液+0.05%Triton】：○

メーカー推奨：WB x5000~10000
ホルマリンは37％ホルマリン液（ナカライ）をPBSやRingerで１/10に希釈
Formalin solution is made by dilution of 37% formalin (Nacalai Inc.) by PBS or Ringer's solution

4%FA in PHEM

Use TBS instead PBS as buffer.

~38 kDa

~38 kDa

4 % PFA fix, PBS+0.05 %Tween20+ 0.5 % TSA block+1 % Donkey serum

【PFA+Triton】:◎,【MeOH】:△,【PFA_70%EtOH】:○【PFA条件】3%PFA/PBS　37℃10min【MeOH条件】MeOH 100% -20℃ 5min【Triton条件】0.1%　Triton-X100/PBS

MDCKII OK.

【4%PFA+0.2%Saponin】:○

3%PFA (in PBS)

0.1% Triton

Golgiが染まります
Stains the Golgi

3%PFA (in PBS)

0.1% Triton

Golgiが染まります
Stains the Golgi

3%PFA (in PBS)

4%PFA (in PBS)

PFA-fixed cryosection/HistoVT one (nacalai) 70˚C 20min

astrocyte, adult neural stem cell

3%PFA (in PBS)

3%PFA (in PBS)

3%PFA (in PBS)

メーカー推奨：WB x5000~10000

4%PFA+0.1%Saponin :○ 0.1%glutalaldehyde + 4% PFA: ○

1%FA

4%PFA (in PBS)

PFA-fixed cryosection/HistoVT one (nacalai) 70˚C 20min

4%PFA (in PBS)

PFA-fixed cryosection/HistoVT one (nacalai) 70˚C 20min

4%PFA (in PBS)

PFA-fixed cryosection/HistoVT one (nacalai) 70˚C 20min

4%PFA (in PBS)

PFA-fixed cryosection/HistoVT one (nacalai) 70˚C 20min

4% PFA

【PFA+Triton】:○,【PFA+Saponin】:○,【IS】:○,【MeOH】:○【PFA条件】3%PFA/PBS　37℃10min【MeOH条件】MeOH 100% -20℃ 5min【Triton条件】0.1%　Triton-X100/PBS【saponin条件】0.05%　saponin /PBS

MDCKII OK.

【PFA+Triton】:◎【PFA条件】3%PFA/PBS　37℃10min【Triton条件】0.1%　Triton-X100/PBS

MDCKII OK.

【Formalin in PBS+0.05%Triton】:○

Alexa Fluor 488 Labeled
ホルマリンは37％ホルマリン液（ナカライ）をPBSやRingerで１/10に希釈
Formalin solution is made by dilution of 37% formalin (Nacalai Inc.) by PBS or Ringer's solution

3%PFA (in PBS), methanol

3%PFA (in PBS)

3%PFA (in PBS)

3%PFA (in PBS), methanol

【PFA+Triton】:◎,【PFA+Saponin】:○【PFA条件】3%PFA/PBS　37℃10min【Triton条件】0.1%　Triton-X100/PBS【saponin条件】0.05%　saponin /PBS

MDCKII OK.

IF: -20℃ methanol or acetone:methanol=1:1

【4%PFA+0.2%Saponin】:○

【4%PFA+0.2%Saponin】:○

4％PFA +0.1％Triton

3%PFA (in PBS)

0.05%saponin

3%PFA in PBS

0.05% saponin

Ab in Golgi sampler kit (BD Transduction Lab)

3%PFA in PBS

0.05% saponin

Ab in Golgi sampler kit (BD Transduction Lab)

4% PFA at 4oC over night

0.5% Triton in PBS

MeOH, PFA

0.05% triton X-100 for PFA fixation

3.7% Formaldehyde (in PBS)

Methanol (-20˚C)

3%PFA (in PBS), methanol

3%PFA (in PBS), methanol

3.7% Formaldehyde (in PBS)

Methanol (-20˚C)

3%PFA (in PBS)

0.05%saponin

3%PFA in PBS

0.05% saponin

Ab in Golgi sampler kit (BD Transduction Lab)

3%PFA in PBS

0.05% saponin

Ab in Golgi sampler kit (BD Transduction Lab)

4%PFA (in phosphate buffer)

0.1% TritonX-100/PBS

4%PFA+0.1%Saponin :○

【PFA+Saponin】:○

交差する動物種：ヒト
【FACS】:○（final conc）2 μg/mL

【PFA+Saponin】:○

交差する動物種：マウス
【FACS】:○（final conc）2 μg/mL

3%PFA (in PB)

acinar細胞のzymogen顆粒が非常に強く染まる。
Zymogen granules are very strongly stained in IF

3%PFA (in PBS), methanol

3%PFA (in PBS), methanol

3%PFA (in PBS), methanol

【PFA+Triton】:×,【PFA+Saponin】:×,【IS】:×,【MeOH】:×【PFA条件】3%PFA/PBS　37℃10min【MeOH条件】MeOH 100% -20℃ 5min【Triton条件】0.1%　Triton-X100/PBS【saponin条件】0.05%　saponin /PBS

4%PFA/PBS,0.01%TritonX100

3%PFA (in PBS)

0.05%saponin

3%PFA (in PBS)

0.05%saponin

3%PFA (in PBS)

0.05%saponin

3%PFA (in PBS)

0.05%saponin

3%PFA (in PBS)

3%PFA in PBS

0.05% saponin

methanol固定でも良く染まる　
Good staining by methanol fix.

3%PFA in PBS

0.05% saponin

Ab in Golgi sampler kit (BD Transduction Lab)